Karamian Roya
Department of Biology, Faculty of Sciences, Bu-Ali Sina University, Hamadan, Iran.
Pak J Biol Sci. 2007 Feb 15;10(4):659-63. doi: 10.3923/pjbs.2007.659.663.
A protocol has been developed for plant regeneration from protoplast culture of Crocus pallasii subsp. haussknechtii using regenerable embryogenic calli obtained from shoot meristem culture on MS+9.28 microM kinetin+4.52 microM 2,4-D. Protoplasts were isolated directly from embryogenic calli, embedded in Ca-alginate beads and cultured with nurse cells in MS+4.64 microM kinetin+4.52 microM 2,4-D+5.68 microM ascorbic acid+0.3 M mannitol at 20 +/- 2 degrees C in darkness. After appearing ofmicrocalli on the surface of the beads, they were transferred onto 1/2MS+2.32 microM kinetin+2.26 microM 2,4-D+5.68 microM ascorbic acid for growth of embryogenic calli. Somatic embryos matured on MS medium growth regulator free and germinated on 1/2MS+14.45 microM GA3 +4.43 microM BA at 20 +/- 2 degrees C in a 16/8 h light/dark cycle.
已开发出一种从番红花属帕拉斯亚种豪氏番红花原生质体培养中再生植株的方案，该方案使用从MS + 9.28微摩尔激动素 + 4.52微摩尔2,4 - D的茎尖分生组织培养获得的可再生胚性愈伤组织。原生质体直接从胚性愈伤组织中分离出来，包埋在钙藻酸盐珠中，并与看护细胞一起在MS + 4.64微摩尔激动素 + 4.52微摩尔2,4 - D + 5.68微摩尔抗坏血酸 + 0.3 M甘露醇中于20±2℃黑暗条件下培养。在珠粒表面出现微愈伤组织后，将它们转移到1/2MS + 2.32微摩尔激动素 + 2.26微摩尔2,4 - D + 5.68微摩尔抗坏血酸上以促进胚性愈伤组织生长。体细胞胚在无生长调节剂的MS培养基上成熟，并在1/2MS + 14.45微摩尔赤霉素 + 4.43微摩尔苄氨基嘌呤上于20±2℃、16/8小时光/暗周期下发芽。
